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bFGF、IGF-Ⅰ对兔关节软骨细胞Ⅱ型胶原表达的影响
Effects of basic fibroblast growth factor and insuline-like growth factor-Ⅰ on the expression of typeⅡ collagen in cultured rabbit articular chondrocytes in vitro
【摘要】 目的:研究碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)、胰岛素样生长因子-Ⅰ(insuline-likegrowthfactor-Ⅰ,IGF-Ⅰ)单独及联合应用对体外培养多次传代的兔关节软骨细胞Ⅱ型胶原表达的影响,探讨传代多次的软骨细胞作为软骨组织工程种子细胞的可行性。方法:第3、5、7代体外培养的兔关节软骨细胞,以常规培养组作为对照组,bFGF(5ng/ml)或和IGF-Ⅰ(100ng/ml)单独刺激及联合刺激作为实验组,免疫组化法测定Ⅱ型胶原的定性表达,并通过图像分析作半定量分析。结果:对照组细胞第5代以后无阳性表达。第3、5、7代细胞在bFGF或IGF-Ⅰ刺激下,Ⅱ型胶原表达强度显著高于对照组(P<0.01);bFGF和IGF-Ⅰ联合作用下,Ⅱ型胶原表达强度均显著高于单独以bFGF或IGF-Ⅰ刺激组(P<0.05)。结论:bFGF、IGF-Ⅰ能促进多次传代软骨细胞Ⅱ型胶原的表达,且联合应用时促进作用更为明显。多次传代软骨细胞在生长因子刺激下,仍有再分化的能力,而有成为软骨组织工程种子细胞的可能。
【Abstract】 Objective: To investigate the effects of single and combined use of basic fibroblast growth factor (bFGF )and insuline-like growth factor-Ⅰ(IGF-Ⅰ)on the expression of typeⅡ collagen in cultured rabbit articular chondrocyte of different passages in vitro. Methods: Cultured chondrocytes of passage 3, 5 and 7 were stimulated by bFGF(5 ng/ml), IGF-Ⅰ(100 ng/ml), bFGF(5 ng/ml)+IGF-Ⅰ(100 ng/ml) respectively. The expression of typeⅡ collagen was tested by immunohistochemistry method qualitatively and analyzed by image analysis semiquantitatively. Results: Regular cultured chondrocytes after passage 5 had no positive expression of typeⅡ collagen. The expression of typeⅡ collagen in the chondrocytes of passage 3, 5 and 7 stimulated by bFGF or IGF-Ⅰwas higher than that of regular cultured group (P < 0.01), and the expression of typeⅡ collagen in bFGF+IGF-Ⅰgroup was significantly higher than that of bFGF or IGF-Ⅰgroup respectively (P < 0.05). Conclusion:bFGF or IGF-Ⅰ promotes typeⅡ collagen expression in multi-passage chondrocytes and the combination use of bFGF and IGF-Ⅰ has synergistic effect. The results suggest that multi-passage chondrocytes stimulated by growth factors are capable to express typeⅡ collagen.
【Key words】 chondrocyte; tissue engineering; basic fibroblast growth factor; insuline-like growth factor-Ⅰ; typeⅡ collagen;
- 【文献出处】 南京医科大学学报(自然科学版) ,Acta Universitatis Medicinalis Nanjing(Natural Science) , 编辑部邮箱 ,2006年05期
- 【分类号】Q813.11
- 【被引频次】10
- 【下载频次】166